See main article: 2000 Italian Open (tennis).
Monica Seles defeated Amélie Mauresmo in the final, 6–2, 7–6(7–4) to win the women's singles tennis title at the 2000 Italian Open. It was her third title of the year, and the 47th of her career.
Venus Williams was the defending champion, but lost in the third round to Jelena Dokic.
The first eight seeds received a bye into the second round.